Efficient Delivery of p53 and Cytochrome C by Supramolecular Assembly of a Dendritic Multi-Domain Delivery System

• Published in: Advanced healthcare materials 2013-12, Vol.2 (12), p.1620-1629
• Main Authors: Ng, David Yuen Wah, Fahrer, Jörg, Wu, Yuzhou, Eisele, Klaus, Kuan, Seah Ling, Barth, Holger, Weil, Tanja
• Format: Article
• Language: English
• Subjects: Apoptosis - drug effects; biohybrids; Biotin - chemistry; Cell Line, Tumor; Cell Survival - drug effects; Conjugation; Cytochromes; Cytochromes c - administration & dosage; Cytochromes c - chemistry; Cytochromes c - pharmacokinetics; Delivery systems; Dendrimers - administration & dosage; Dendrimers - chemistry; Dendrimers - pharmacokinetics; dendronized proteins; HeLa Cells; Humans; Medical research; Nanostructure; p53 and cytochrome c delivery; Proteins; Self assembly; Streptavidin - chemistry; streptavidin adapters; supramolecular drugs; Tumor Suppressor Protein p53 - administration & dosage; Tumor Suppressor Protein p53 - chemistry; Tumor Suppressor Protein p53 - pharmacokinetics; Uptakes; Viability
• ISSN: 2192-2640; 2192-2659
• DOI: 10.1002/adhm.201200419

Versatile nanocarrier systems facilitating uptake of exogenous proteins are highly alluring in evaluating these proteins for therapeutic applications. The self‐assembly of an efficient nano‐sized protein transporter consisting of three different entities is presented: A streptavidin protein core functioning as an adapter, second generation polyamidoamine dendrons for facilitating cell uptake as well as two different therapeutic proteins (tumor suppressor p53 or pro‐apoptotic cytochrome c as cargo). Well‐defined dendrons containing a biotin core are prepared and display no cytotoxic behavior upon conjugation to streptavidin. The integration of biotinylated human recombinant p53 (B‐p53) into the three component system allows excellent internalization into HeLa, A549 and SaOS osteosarcoma cells monitored via confocal microscopy, immunoblot analysis and co‐localization studies. In addition, the conjugation of B‐p53 to dendronized streptavidin preserves its specific DNA‐binding in vitro, and its delivery into SaOS cells impairs cell viability with concomitant activation of caspases 3 and 7. The versatility of this system is further exhibited by the significant enhancement of the pro‐apoptotic effects of internalized cytochrome c which is analyzed by flow cytometry and cell viability assays. These results demonstrate that the “bio‐click” self‐assembly of biotinylated dendrons and proteins on a streptavidin adapter yields a stable supramolecular complex. This efficient bionanotransporter provides an attractive platform for mediating the delivery of functional proteins of interest into living mammalian cells in a facile and rapid way. A versatile and efficient multidomain delivery system facilitating the cell uptake of the proteins p53 and Cytochrome C is obtained by supramolecular assembly. Such nanotransporters contain a streptavidin protein core functioning as a bio‐adapter, second generation polyamidoamine dendrons for cell uptake as well as a therapeutic protein as cargo. The delivery and functional activity of tumor suppressor p53 or pro‐apoptotic cytochrome c are studied in detail.